6beta, 17alpha, 21-trihydroxy-4-pregnene-3,20-dione and esters thereof



UNITED STATES PATENT OFFICE 65,17m21-TRlHYDRoXY- l-PREGNENE -iififl- DIONE ESTERS THEREOF Herbert 0. Murray, Hickory Corners, and Darcy H. Peterson, Kalamazoo, Mich., assignors to The Upjohn Company, Kalamazoo, Mich.,. a corporation of Michigan No Drawing. Application July I, 1952, S ri l No- 296,735

15 Claims. (Cl. 260-39747.)

2 This invention relates to new and useful comby reacting 6p,170;,21trihydroxy-4-preanenepounds and more specifically to oel'hzl-tri- 3,20-dione with ketene, ketenes of selected acids, hydroxy-4-pregnene-3,26:dione and the carselected acids, acid anhydrides or acid chlorides, box-ylic acid esters of 6p,1'7c,21-trihydroXy-4- in an organic solvent such as pyridine or the pregnene-3,20-dione and to methods for the prep- 5 like. Representative 21-mono and 6,21 di-fisliers aration thereof. thus prepared include, among others, one to eight It is an object of this invention to provide the carbon atom carhoxylic acid acyloxy esters of novel 6p,17u,2l-trihydrox-y-4-pregnene-3-,26-dione saturated or unsaturated, aliphatic or carbo. and the monoand di-esters of 6,6,17m2l-tricyclic, cycloaliphatic, aryl, arylalkyl, alkaryl, hydroxy-4-pregnene-3,ZO-dione having phar 10 mono-, dior polycarboxylic acids which form rnaoological and especially progestational activity ester groups such as, for example, formoylo cy, and beingadditionally useful in the synthesis acetoxy, propionyloxy, butyryloxy, valeryloxy, of various oxygenated steroids. Another object hexanoyloxy,heptanoyloxy, qclianoyloxmbenzoxy, is to provide a process for the preparation of these phenylacetoxy, toluoyloxy, naphthoyloxy, cycle, new and useful compositions of matter. Other pentylformoyloxy,,8-cyclopentylpropionyloxy,ac!-

objects of the invention will be apparent to those rylylcxy, eyclohe xyl-formy-loxy, the half and die skilled in the art to which this invention pertains. esters of malonic, maleic, sucoinio, glutaric and The novel compounds of the present invention adipic acids, and the like. The. acids may also are represented by the following structural forcontain non-interfering substituents, such as mula: monoor po y-h o chlo a bronze. hydroxv.

m t xy, and the like, if desired. I

Th startin material in these acylating reactions, 6,6,17u,21 e trihydroxyl-pregnene 3,20; dione, may be prepared by subjecting ll-desoxyzl7-hydroxyoorticosterone to the action of; a fungus of the order Muoora1es as set, forth, in the applications of Murray and Peterson, Serial Number 180,4:96, filed August 19, 1.950, new ahan doned, Serial Number 272,944, filed, February 2-3;, 2; ssued on. Jul 5 a U ted S a Pa ent 2,602 769, and Ser al Number 2931651, filed June 14 1952, of which this application is a continuation-impart.

The following preparation and examples are illustrative of the objects, processes; and products of the present invention, but are not to be construed as limiting.

wherein R, is a member selected from, the group consisting of hydrogen and an acyl group of a carxy ic ac d These compounds are useful as intermediates in the synthesis of pharmacologically active steroids. For example, on oxidation of 613,21- mple 1.- 6 ',3,17a,21-trzhydroxy-4-pregnenediacetoxy-1la-hydroxy-4-pregnene-3,20-dione to g gmdi 6,6-acetoxy-4-androstene-3,17dione, followed by pyrolysis of this compound under reduced pres- A. medium was, prepared of twenty grams of sure to obtain 4,6-androstadiene-3,17Qdione and Edamine enzymatic digest of lactalbu-rnin, three then hydrogenation of this product by one mole grams of corn, steep 111. 1 and if y grams Oi of hydrogen in the presence of a charcoal-pale hn cal dextrose diluted to one liter with ill? ladium catalyst, the androgenically active steroid, wa er and: adjusted to a pH of 4.3 to- 4,.5. Two 4-an drostene-3,17-dione is obtained. This. comliters o th s sterilized medium W 5 n ulat d pound may be converted to testosterone by the with Rhz'eopus ar h i A 1145,, and in: action of fermenting yeast. cubatedfor 24 hours at a temperature of 23 In addition, 6 3,1'Za,2l-trihyd-roxyl-pregnenedegrees centigrade using; rate. of aerat on and 3,20-dione and esters thereof demonstrate cortistirring such that the oxygen uptakewas 63 cold, androgenic, progesterone, anabolic, antito 7 millimoles per hour per liter of NazSQa hypertensive, and anesthetic properties. according to the method of Cooper, Fernstrom Using the procedure set forth in the following and Miller, Ind. Eng Chem, 3.5, 564 1944:) 13o examples, the monoand di-esters of 6p,1'7a,21- this, medium containing a 24-hour growthor trihydroxy-l-pregnene-3,20-dione are prepared Rhizome orrltiaus; was. added one gram of 11;

3 desoxy-l'l-hydroxycorticosterone in milliliters of acetone to provide a suspension of the steroid in the culture. After an additional 42-hour period of incubation under the same conditions of temperature and aeration, the beer and mycelium were extracted. The mycelium was filtered, washed twice, each time with a volume of acetone approximately equal to the volume of the mycelium and extracted twice, each time with a volume of methylene chloride approximately equal to the volume of the mycelium. The acetone and methylene chloride extracts, including solvent, were added to the beer filtrate. The mixed extracts and beer filtrate were extracted successively with two one-half by volume portions of methylene chloride and then with two one-fourth by volume portions of methylene chloride. The combined methylene chloride extracts were washed with two one-tenths by volume portions of a two percent aqueous solution of sodium bicarbonate and then with two one-' tenth by volume portions of water. After drying the methylene chloride with about three to five grams of anhydrous sodium sulfate per liter of solvent and filtering, the solvent was removed by distillation. The residue was dissolved in a minimum of methylene chloride, filtered and the solvent was then evaporated in air or on a steam bath. The resulting crude crystals were dried and then washed five times portions of ether per gram of crude crystals weighing 1.834 grams. Extraction of the crystals with three portions, five milliliters each. of ice cold methylene chloride yielded 525 milligrams of crystals melting at 198 to 20? degrees centigrade. Recrystallization from a mixture of thirteen milliliters of methanol and one milliliter of water produced 305 milligrams of crystals, melting at 224 to 228 degrees centigrade. Recrystallization from methanol of milligrams yielded 26 milligrams of crystals, 6p,l7a,2l-trihydroxy- 4-pregnene-3,20-dione, melting at 224 to 22'? degrees centigrade, having an optical rotation of [0.1 of plus 58.5 degrees (0.894 in percent ethanol). Infrared microcombustion and acetylation indicated the addition of one hydroxyl group.

Analysis: Calculated for (3211-13005: C, 69.58; H, 8.35. Found: C, 69.66; H, 8.22.

The identical procedure using organism Rhizozms nigricans minus strain ATCC 6227b instead of Rhieopus arrhz'eus similarly produced compound 648,1Ta,21 trihydroxyl-pregnene 3,20- dione from compound 8., il-desoxy-l'I-hydroxycorticosterone.

Example 2.--6p,17a-Dihydromy-21-acetoxy-4- pregnene-3,20-dione A solution containing 0.031 milliliter of redistilled acetic anhydride dissolved in two milliliters of pyridine was added to a solution containing milligrams of 6p,17a,21-trihydroxy-4-pregnone-3,20-dione dissolved in one milliliter of pyridine. After allowing the mixture to stand at room temperature for sixteen hours, forty milliliters of water was added thereto. Within a short time, crystals began to form. ihe reaction mixture was then refrigerated for three hours, centrifuged, washed with water and dried under vacuum. A yield was obtained of 49.5 milligrams of crystalline 6p,1'7a-dihydroxy-21- acetoxy-4-pregnene-3,20-dione, melting at 246 to 260 degrees centigrade. Infrared analysis verifled this compound as the 2l-acetate since there is a 1:1:1 ratio of acetate: normal ketone: conwith five-milliliter jugated ketone. Recrystallization from 2.8 milliliters of methanol yielded twenty milligrams of the purified compound melting at 256 to 265 degrees centigrade.

Example 3.-6a., 21 diacetomy, 17 hydroxy 4- pregnene-3,20-dione A sample of 51.1 milligrams of 6fi,17a,21t1i hydroxy-4-pregnene-3,20-dione was dissolved in four milliliters of acetic anhydride and two milliliters of pyridine. The solution was warmed for a short time on a steam bath to dissolve the crystals completely. After maintaining the mixture for fifteen hours at room temperature, it was diluted with thirty milliliters of ice water and extracted twice with thirty milliliter portions of a mixture of five parts of ether to one part of methylene chloride. The combined extracts were washed twice with ten milliliter portions of 2 N hydrochloric acid, once with a ten milliliter portion of water, three times with ten milliliter portions of seven percent sodium bicarbonate, three times with ten milliliter portions of water and the solvents dried over anhydrous sodium sulfate. After removal of the solvents in vacuo, 58.5 milligrams of crystalline residue remained. Recrystallization from five milliliters of acetone by the dropwise addition of ether yielded 34 milligrams of crystalline 6,8,21- diacetoxy,17a-hydroxy-4 pregnene 3,20 dione melting at 192 to 195 degrees centigrade, [11],, of plus 63 degrees (0.9852 in chloroform).

Analysis: Percent calculated for (325K340?! C, 67.24; H, 7.67. Found: C, 67.30; H, 7.68.

Although the acylation processes illustrated by Examples 1 and 2 produce both the mono-esters and the di-esters, the proportions of the esters obtained in this manner depend upon the proportions of acylating agent to 6p,1'7a,21-trihydroxy-4-pregnene-3,20-dione. By using one mol of 6p,l7a,21 trihydroxy 4 pregnene-3,20-dione with about one mol or equivalent of acylating agent, the mono-acylated compound is the predominant product while with the use of about two mols or equivalents of acylating agent, the diacylated compound is the predominant product.

Example 4.--6;8,17a-dihyd1ory,21 formylomy-4- pregnene-3,20-dione Example 5.-6B,21-diformyZo:cy-17a-hydroxy-4- pregnene-3,20-dione Following the procedure of Example 4 and using a solution containing 0.5 gram of 6fi,1'7a,21- trihydroxy-4-pregnene-3,20-dione dissolved in twenty milliliters of formic acid, crystals of 65,21 diformloxy 17a hydroxy 4 pregnene- 3,20-dione were produced.

Ewample 6.-6e,17s-dihydro:cy-21 propionyloscy- 4-pregnene-3,20-dione Following the procedure of Example 2 and using an equivalent proportion of propionic anhydride in place of acetic anhydride, crystals of 6e,17edihydroxy-2l-propionyloxy 4 pregnene- 3,20-di0ne were produced.

5 Example 7.6;9,21-dipropionyZoxy-17a 4-prcgnene-3,20-dione Following the procedure of Example 3 and using an equivalent proportion of propionic anhydride in place of acetic anhydride, 6,8,21-dipropionyloxy-17a-hydroxy-4 pregnene 3,20-dione was produced.

Eazample 8.65,1 7u-dihydromy-21 -benz0a:y- 4-pregnene--3,20-dione To a solution containing 0.2 gram of 65, 1711,21- trihydroxy-4-pregnene-3,20-dione dissolved in eight milliliters of benzene was added 0.4 milliliter of freshly dried and redistilled pyridine and 0.2 milliliter of freshly distilled benzoyl chloride and the mixture was maintained at room temperature for twenty hours. The reaction mixture was then diluted with fifty milliliters of ether, washed successively with water, ten percent aqueous sodium hydroxide solution, and again with water, dried over anhydrous sodium sulfate, filtered and then evaporated free of solvent. The residue Was mixed with 25 milliliters of water and maintained under reflux for fifteen hydroxyminutes, cooled, extracted with ether, washed with cent water followed by a wash with a ten persodium hydroxide solution, and again washed with water, dried over anhydrous sodium sulfate, filtered and evaporated to a volume of ten milliliters. After cooling, the resulting crystals were removed by filtration and washed with benzene to yield crystalline 6,8,17a-dihYdI'0XY-21- benzoxyl-pregnene-3,20-dione.

Example 9.6p,21 -d2'benzoa:z -1 7e-hydroxy- 4-pregnene-3,20-dione Following the procedure of Example 8 and increasing the quantity of benzoyl chloride to 0.4 milliliter, 65,21-dibenzoxy-17a-hydroxy4-pregnene-3,20-dione was produced.

If a mixed ester containing two dissimilar acyl groups is desired, 6B,17,21-trihydroxy-4-preg nene-3,20-dione may be partially esterified with one acylating agent and the resulting monoester may then be completely esterified with another acylating agent ferent acyl group.

It is to be understood that the invention is not to be limited to the exact details of operation or exact compounds shown and described herein, as obvious modifications and equivalents will be apparent to one skilled in the art, and the invention is therefore to be limited only by the scope of the appended claims.

We claim:

1. A compound selected from the group con sisting of 6,8,l7e,21-trihydroxyl-pregnene-3,20- dione and 6,6,17a,21-trihydroxy-4-pregnene-3,20- dione esters of hydrocarbon carboxylic acid containing less than nine carbon atoms.

2. 6,3,17a,21 trihydroxy 4 pregnene 3,20-dione.

3. 6,9,17a,21 trihydroxy 4 pregnene 3,20 dione esters of hydrocarbon carboxylic acid containing less than nine carbon atoms.

4. 6,8,17a dihydroxy 21 acetoxy 4 pregnene-3,20-dione.

which introduces a dif- 5. 65,21 diacetoxy '17-12 hydroxy 4 pregnene-3,20-dione.

6. 6 8,174; dihydroxy 21 pregnene-3,20-dione.

7. 63,21 dipropionyloxy 17a pregnene-3,20-dione.

8. A process for the preparation of esters of 5,8,17a,21 trihydroxy 4 pregnene 3,20 diones which comprises reacting 6p,17a,21-trihydroxy-4-pregnene-3,20-diones with an acylating agent to produce esters of 6,8,17a,21-trihydroxy-4-pregnene-3,20-dimes.

9. A process for the preparation of esters o1 6fi,17a,21 trihydroxy 4 pregnene 3,20 dione which comprises reacting 6fl,17a,21-trihydroxy-4-pregnene-3,20-diones with a carboxylic acid to produce esters of- 6/3,17u,21-trihydroxy- 4-pregnene-3,20-diones.

10. A proces for the preparation of esters of 6p,17a,21 trihydroxy 4 pregnene 3,20 diones which comprises reacting 6fi,17a,21-trihydroxy-4-pregnene-3,20-diones with a carboxylic acid anhydride to produce esters of 6,8,l'7a,21-trihydroxy-4-pregnene-3,20-diones.

11. A process for the preparation of esters of 6B,17a,21 trihydroxy 4 pregnene 3,20 diones which comprises reacting 6,8,17a,21-trihydroxy-4-pregnene-3,20-diones with a carboxylic acid halide to produce esters of 6;8,17a,21- trihydroxy-4-pregnene-3,20-diones.

12. A process for the preparation of 6/3,17a-dihydroxy 21 acyloxy 4 pregnene 3,20 diones which comprises reacting equivalent amounts of 6p,17a,21-trihydroXy4-pregnene- 3,20-diones with an acylating agent to produce 6 8,1'h-dihydroxy-21-acyloxy-4-pregnenes.

13. A process for the preparation of 6,6,21-diacyloxy 17a hydroxy 4 pregnene 3,20 diones which comprises reacting 6j3,17a,21-t1ihydroxy-4-pregnene-3,20-diones with an acylating agent in a ratio of about one mol to about two mols, respectively, to produce 6,6,21-diacyloxy-l7a-hydroxy-4-pregnene-3,20-diones.

14. A process for the preparation of 6,6,17a-dihydroxy 21 acetoxy 4 pregnene 3,20 dione which comprises reacting equivalent amounts of 6 3,17)1-trihydroxy-4-pregnene- 3,20-dione with acetic anhydride to produce 6fi,17a dihydroxy 21 acetoxy 4 pregnene 3,20-dione.

15. A process for the preparation of 6,6,21-diacetoxy 17a hydroxy 4 pregnene 3,20 dione which comprises reacting 6]3,17oa,21-12ri hydroxy-4-pregnene-3,20-dione with acetic anpropionyloxy 4 hydroxy 4 hydride in a ratio of about one mol to about two mols, respectively, to produce 6,3,21-diacetoxy-17a-4-pregnene-3,ZO-dione.

HERBERT C. MURRAY. DUREY H. PETERSON.

References Cited in the file of this patent UNITED STATES PATENTS Number Name Date 2,337,584 Marker Dec. 28, 1943 2,352,848 Marker July 4, 1944 2,366,204 Marker Jan. 2, 1945 2,423,517 Reichstein July 8, 1947 

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF 6B,17A,21-TRIHYDROXY-4-PREGNENE-3,20DIONE AND 6B,17A,21-TRIHYDROXY-4-PREGNENE-3,20DIONE ESTERS OF HYDROCARBON CARBOXYLIC ACID CONTAINING LESS THAN NINE CARBON ATOMS. 